Effect of protein malnutrition on the crosslinking of dermal collagen in aging.

The effect of protein malnutrition on collagen crosslinking was studied in different age groups of rats. The percent of reversibility of neutral salt-soluble collagen gel and the susceptibility of insoluble collagen to denaturing agents and pronase decreased with aging. The electrophoretic pattern of neutral salt-soluble collagen on sodium dodecyl sulphate polyacrylamide gels showed a decrease of alpha 1 and alpha 2 subunits and an increase of beta-components, resulting in a decrease of alpha/beta ratio in aging. There was a significant decrease in the aldehyde content of neutral salt-soluble collagen during age. Protein deficiency was found to impair the crosslinking and the effect was greater in younger animals than in older ones.

Glycoprotein nature of tannase in Aspergillus niger.

1. Tannase (tannin acyl-hydrolase, EC 3.1.1.20) was isolated from the culture medium of Aspergillus niger and purified about 200-fold. On polyacrylamide-gel electrophoresis it gave a single band. 2. The molecular weight of the enzyme was of the order of 55 000 as determined by gel filtration. The enzyme contains 21.5% of carbohydrates (mannose and glucose). 3. Treatment of tannase with alkaline borohydride decreased the content of threonine, serine and mannose, suggesting that the carbohydrate-peptide linkage is of the O-glycoside type, involving mannose linked to threonine and serine.

Hydroxyproline-rich glycoprotein in the seed coat of field bean.

1. The seed coats of field bean (Dolichos lablab var. lignosus) were found to contain bound hydroxyproline (0.26 g per 100 g dry weight) extractable in 30% with 5% trichloroacetic acid or in 90% with 10% potassium hydroxide. 2. The alkaline extract was fractionated by DEAE-cellulose chromatography into several hydroxyproline-containing fractions, composed of protein and carbohydrate in varying proportions. From the major fraction, a hydroxyproline-containing glycoprotein fraction was isolated by Sephadex G-200 chromatography. It gave a single band on agar-gel electrophoresis, and contained: protein (23.1%), arabinose (11.2%), galactose (13.4%), glucose (17.6%), mannose (22.6%) and uronic acids (11.3%). The content of hydroxyproline in the protein moiety was about 9%. Hydrolysis of the glycoprotein fraction with barium hydroxide yielded three components containing hydroxyproline and arabinose at the ratios of 1:2, 1:3 and 1:4.

Effect of protein deficiency on the metabolism of glycoproteins and glycosaminoglycans in albino rat skin.

1. The total content of neutral sugars in skin of the weanling albino rats kept on the protein-deficient diet was increased by about 40%; this was mainly due to the increased concentration of galactose. The content of sialic acid was increased by about 20%. The collagen nitrogen was decreased significantly, with a concomitant increase of non-collagen nitrogen. At the same time, the content of sulphated glycosaminoglycans in skin was significantly decreased and that of non-sulphated glycosaminoglycans was increased. 2. Protein-deficient diet enhanced the activities of the protein-bound carbohydrate-degrading lysosomal hydrolases, viz. cathepsin D (EC 3.4.4.23), N-acetyl-beta-D-glucosaminidase (EC 3.2.1.30) and beta-D-glucuronidase (EC 3.2.1.31) both in liver and skin. The activity of liver hyaluronidase (EC 3.2.1.35) was also increased upon limitation of protein supply. 3. The changes observed in skin were accompanied by increased concentration of the protein-bound hexoses, hexosamines and sialic acids in serum, and of hexosamine and uronic acid in urine. The serum fucose remained unchanged.

Serum beta-N-acetylhexosaminidase, carcinoembryonic antigen & sialic acid in benign & malignant extrahepatic biliary obstruction.

Study of tumour markers to differentiate benign and malignant extra hepatic biliary obstructions showed that levels fo serum beta-N-acetylhexosaminidase and lipid associated sialic acid were not different in the two groups. The levels of carcinoembryonic antigen (CEA) and total sialic acid (TSA), on the other hand, were significantly higher in patients with the malignant biliary obstruction. At a cut-off value of 6.4 ng/ml (mean + 2SD) CEA had 66.7 per cent true positivity in malignancy 100 per cent true negativity in control and 78 per cent true negativity in the benign group. Similarly, TSA at a cut off value of 60 mg/dl had 61 per cent true positivity in malignancy, 90 per cent true negativity in controls and 70 per cent true negativity in the benign group. These two tumour markers appear to have similar diagnostic potential for malignant extrahepatic biliary obstruction. Surgical management of the malignant obstruction did not result in a decline in the elevated levels of these two markers during the post-surgical period of 7 days.

Glycohydrolases and lysosomal stability in adjuvant induced arthritis.

Adjuvant induced arthritis in rats was studied by the changes in serum and urinary protein-bound carbohydrate metabolites, changes in serum and tissue lysosomal glycohydrolases and lysosomal fragility. From the second week onwards the urinary excretion of hexosamine and uronic acid is increased. Serum levels of protein bound hexose, hexosamine, sialic acid and fucose are increased significantly in both the acute and chronic phases of the disease. There is no change in the total activity of lysosomal glycohydrolases, viz., beta-glucuronidase, beta-N-acetyl glucosaminidase and cathepsin D in the tissues of liver, kidney and spleen except that of liver enzymes in the chronic phase which are elevated significantly. The free activities of lysosomal glycohydrolases investigated, viz., beta-glucuronidase, beta-N-acetyl glucosaminidase, beta-galactosidase, alpha-mannosidase and cathepsin D are increased in liver and spleen in the acute phase. The free activities of beta-glucuronidase, beta-N-acetyl glucosaminidase and cathepsin D of kidney showed no change whereas those of beta-galactosidase and alpha-mannosidase are increased. In the chronic phase of the disease the free activities of all glycohydrolases are significantly increased in all tissues. Serum glycohydrolases are significantly increased in both acute and chronic phases. Studies on lysosomal preparations showed increased fragility of lysosomes derived from liver and kidney of arthritic rats in both phases of the disease.

Effect of indomethacin and naproxen on collagen cross-linking.

The influence of indomethacin and naproxen on rat dermal collagen cross-linking was investigated in vivo. The analyses of skins showed that the naproxen group had a decrease in percent reversibility of neutral salt-soluble collagen gel, susceptibility of insoluble collagen to denaturing agents and pronase and alpha/beta ratio of neutral salt-soluble collagen whereas the aldehyde content was significantly increased. Indomethacin was ineffective for all these parameters. The results indicated that both the inter and intra-molecular cross-linking of collagen was increased in the naproxen treated group while indomethacin did not influence the collagen cross-linking.

Effect of adjuvant-arthritis on collagen metabolism.

In albino rats with adjuvant arthritis, the metabolism of skin collagen was studied. This skin samples were taken from 21 to 49 days after the injection of Freund's adjuvant. Glycine-1-C14 was given 5h before sacrifice of the animals. The analyses of skins showed that compared to uninflammed skins, the adjuvant arthritis group had an increase in neutral-salt soluble collagen and acid soluble collagen contents. But the total and insoluble collagen contents were found to be decreased. The incorporation of C14 glycine into skin collagen and the free glycine content of skins were also decreased. There was no significant change in the total nitrogen, RNA and DNA contents of skins. The analyses of urine at weekly intervals showed an increased value of urinary total, free and non-dialysable hydroxyproline in arthritic group. The results suggest that there is an alteration in the metabolism of collagen.

Isolation, characterization and nature of carbohydrate-peptide linkage of an alkali-extractable non-collagenous glycoprotein from albino rat skins.

A glycoprotein was isolated from young albino rat skins by alkali extraction under mild conditions and purified by Sephadex G-200 and DEAE-Sephadex A-50 chromatography. It was found to be homogeneous by agar gel electrophoresis. It had a molecular weight of approximately 90,000 and contained galactose, mannose, fucose, N-acetylglucosamine, N-acetylgalactosamine and sialic acids as its carbohydrate constituents. The release of sialic acids from the glycoprotein by neuraminidase indicated their terminal positions in the carbohydrate chains. The glycoprotein lacked hydroxyproline which indicates its non-collagenous nature. The treatment of the glycoprotein with alkaline borohydride resulted in the decrease of threonine, serine and N-acetylgalactosamine contents. The presence of O-glycosidic linkage of N-acetylgalactosamine with serine and threonine is therefore suggested.

Pericholecystic adhesions in single v multiple gallstones and their consequences for laparoscopic cholecystectomy.

BACKGROUND: Adhesions are amongst the common reasons for open conversion of laparoscopic cholecystectomy. It is not clear whether this problem is more common with single or multiple gallstones. PATIENTS AND METHODS: The clinical records of 110 patients with chronic cholelithiasis harboring multiple stones in the gallbladder (multiple stone group; MSG) and 45 patients with single stones in the gallbladder (SSG) undergoing laparoscopic cholecystectomy were analyzed for differences in the clinical presentation and outcome with special reference to the incidence of pericholecystic adhesions, size of the stones, and their implications for conversion and complications. RESULTS: Patients in the SSG had a significantly higher incidence of dense pericholecystic adhesions in the region of the porta hepatis (P = 0.003). Eleven patients in the SSG (24.4%) were converted to open cholecystectomy. Dense pericholecystic adhesions around the porta hepatis alone contributed to nine of these conversions (81.8%). The size of the stones was significantly greater (P < 0.001) in those patients of the SSG who required conversion to open cholecystectomy. Thirteen patients of the MSG (11.8%) required conversion to open cholecystectomy. Dense pericholecystic adhesions alone contributed to conversion in four cases (30.7%), and the size of the gallstones was not significantly different (P = 0.981) in patients with or without conversion to open cholecystectomy. There was no difference in the clinical presentation or complications in the two groups of patients. CONCLUSION: Dense adhesions in the porta hepatis are significantly more common in patients with single stones and are the most common reason for open conversion.

Anomalous bile duct injury during laparoscopic cholecystectomy.

We report a patient in whom the common hepatic duct drained into the gall bladder body and the cystic duct continued as the bile duct into the duodenum. The anomalous duct was inadvertently injured during laparoscopic cholecystectomy. The injury was repaired and end-to-end anastomosis of the hepatic and cystic ducts was done through a subcostal incision.

Intestinal endometriosis.

Two cases with intestinal endometriosis, one with a right iliac fossa mass and the other with subacute intestinal obstruction, are presented. In both the cases, the diagnosis was not suspected clinically and was reached only on histological examination of the resected specimens.

Effect of blockade of transdiaphragmatic absorption of bacteria on survival in peritonitis: an experimental study in rats.

An experimental rat model of established peritonitis was used to test the effect of intraperitoneal injection of platelet rich plasma (PRP) on blood and peritoneal fluid culture positivity and survival rates. Thirty animals divided into two groups of 15 each were studied. The first group served as control while animals in the second group received intraperitoneal injection of PRP. The use of PRP in established. Peritonitis was of no significant benefit.

Giant perforations of duodenal ulcer.

Over a period of eleven years, eight patients were treated for duodenal ulcer perforation measuring more than one centimetre in diameter. In five of these patients, the perforation was sealed using a jejunal loop as serosal onlay patch; one patient underwent gastrectomy and in two patients catheter duodenostomy was done. Two patients died, both due to renal failure. The six surviving patients were discharged with the advice for a subsequent 'ulcer-curing' operation after six months. Two patients developed life threatening complications during this waiting period and one of these died. Giant perforations of duodenal ulcer can safely be closed using a jejunal loop as serosal patch. Delay in doing the second stage definitive surgery for the ulcer may be dangerous.

Adjuvant interstitial brachytherapy in a case of anorectal melanoma.

PURPOSE: A locally resectable case of anorectal melanoma is reported. In order to prevent local recurrence, interstitial brachytherapy was used. METHOD: A 45 years old emaciated female presented with a 8 x 6 x 6 cm, hard, mobile, intraluminal mass in the anal canal and rectum, biopsy revealed malignant melanoma. As the mass was locally resectable, wide local excision of the tumour was carried out. She was subsequently given interstitial brachytherapy with Caesium--137 implants at tumour bed, not described so far in literature. Six months later her general condition had improved but abdomino-perineal resection was necessitated due to presence of extra-rectal metastasis. RESULTS: Patient is in our follow-up for 36 months now and doing well. CONCLUSION: Supplementation of interstitial brachytherapy after local resection of ano-rectal melanoma may help to prevent local recurrence.

Biliary mishaps in laparoscopic cholecystectomy.

AIM: To determine the incidence and types of biliary complications following laparoscopic cholecystectomy in our patients. METHODS: The clinical records of one hundred and fifty-five patients undergoing laparoscopic cholecystectomy were reviewed. RESULTS: Five patients developed biliary mishaps. The overall incidence of biliary complications was 3.2% (5/155). The incidence of major complications was 1.9% (3/155) and the incidence of minor biliary complications was 1.2% (2/155). In 3 out of 5 patients the mishap was attributed to developmental anomalies. Dense pericholecystic adhesions and cystic duct blow out were responsible for biliary complications in one patient each. Bilioenteric anastomosis was performed in two patients and restoration of continuity of the common hepatic duct over a T-tube was done in one patient. Side hole in an accessory duct was repaired over a T-tube and cystic duct blow out was managed with endoscopic biliary drainage alone. CONCLUSIONS: A high index of suspicion of developmental anomalies, cautions approach in difficult cases and readiness to consider conversion to open cholecystectomy are recommended to reduce the incidence of biliary complications in laparoscopic cholecystectomy.

Clinical significance of carcinoembryonic antigen in colorectal malignancy.

Carcinoembryonic antigen (CEA) assay was performed in 40 patients of histologically proven colorectal carcinoma. The overall incidence of positivity was 72.5%. The incidence increased from 40% in Duke's A stage to 84.6% in Duke's C stage. Similarly the mean CEa levels also increased as the disease advanced i.e. 4.96 ng/ml, 8.07 ng/ml and 12.7 ng/ml in Duke's A, B and C respectively. Cancer with poor prognosis i.e. poorly differentiated and colloid carcinoma, had significantly less rise in CEA values (P < 0.05) as compared to well differentiated carcinoma. There was no relation of CEA values with the gross appearance of the tumour and lymph node involvement. CEA level came down in all the patients after surgery. Based on the postoperative CEA estimation, complete tumour clearance had been achieved in 86.2% of patients.

Management of duodenal trauma.

The surgical management of fifteen cases of duodenal trauma is reported. Blunt injury of the abdomen was the commonest mode of injury. Thirteen patients (86.6%) had associated intra-abdominal and/or extra-abdominal injuries. Diagnosis of duodenal injury was made at laparotomy in all patients. In the majority of patients (n = 11), either duodenal repair, or anastomosis with the pylorus or jejunum was done. Duodenal diverticulization or pancreaticoduodenectomy were undertaken in 2 patients each. Feeding jejunostomy was an important adjunct for nutritional support. Septicemia with multisystem organ failure and duodenal fistula were lethal complications. Mortality in the present series was 46.6%.

Isolated splenic vein thrombosis with natural shunt caused by jejunal tuberculosis.

A case of jejunal tuberculosis causing isolated splenic vein thrombosis with natural shunt is reported, for the first time. The patient, presented with upper small bowel obstruction and massive upper gastrointestinal bleeding, was treated successfully with splenectomy, jejunal resection and antituberculous drugs.

Multiple neoplasia in a patient with pituitary acromegaly.

A case of pituitary acromegaly with multiple nonendocrine neoplasia is reported. The tumorogenic potential of growth hormone, prolactin and other growth factors is discussed.